Sequencing Reaction Purification—NucleoSEQ

Unincorporated dye terminators negatively affect the analysis of sequencing results: an excess of dye terminators causes so-called "dye blobs", which result in a partly unreadable sequence.

NucleoSEQ removes unincorporated dye terminators, so the subsequent analysis is of high quality with long read length and minimized background. NucleoSEQ columns are designed for fast, effective, and cost efficient sequencing reaction clean-up. The spin columns are prefilled with a dry size-exclusion matrix for efficient removal of dye terminators (e.g., BigDye terminators). The gel-filtration material consists of spheres with uniform pores, and separates molecules according to molecular weight. After applying the seqencing reaction to the NucleoSEQ column, the small dye terminators and other impurities e.g. salts, nucleotides, primers, and traces of organic solvents are retained in the pores while labeled DNA fragments are excluded and recovered in the flowthrough with high yield.

In order to achieve long-term storage at room temperature, NucleoSEQ columns are prefilled with dry gel filtration resin. The matrix can easily be hydrated by adding water followed by an incubation period (>30 min). Hydrated columns are ready-to-use and can be stored at 4°C for 14 days. A short centrifugation step removes the remaining storage buffer. After loading the sample onto the column followed by a second centrifugation step, the DNA fragments of interest are recovered in the flow-through.